RM2235. Rotary Microtome. Instructions for Use

Transcription

1 Instructions for Use RM2235 Rotary Microtome Leica RM2235 V 2.1, English 11/2015 Order No.: RevF Always keep this manual with the instrument. Read carefully before working with the instrument.

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3 NOTE The information, numerical data, notes and value judgments contained in this manual represent the current state of scientific knowledge and state-of-the-art technology as we understand it following thorough investigation in this field. We are under no obligation to update the present manual periodically and on an ongoing basis according to the latest technical developments, nor to provide our customers with additional copies, updates etc. of this manual. To the extent permitted in accordance with the national legal system as applicable in each individual case, we shall not be held liable for erroneous statements, drawings, technical illustrations etc. contained in this manual. In particular, no liability whatsoever is accepted for any financial loss or consequential damage caused by or related to compliance with statements or other information in this manual. Statements, drawings, illustrations and other information as regards contents or technical details of the present Instructions for Use are not to be considered as warranted characteristics of our products. These are determined only by the contract provisions agreed between ourselves and our customers. Leica reserves the right to change technical specifications as well as manufacturing processes without prior notice. Only in this way is it possible to continuously improve the technology and manufacturing techniques used in our products. This document is protected under copyright laws. All copyrights to this documentation are held by Leica Biosystems Nussloch GmbH. Any reproduction of text and illustrations (or of any parts thereof) by means of print, photocopy, microfiche, web cam or other methods including any electronic systems and media requires express prior permission in writing by Leica Biosystems Nussloch GmbH. For the instrument serial number and year of manufacture, please refer to the nameplate on the back of the instrument. Leica Biosystems Nussloch GmbH Leica Biosystems Nussloch GmbH Heidelberger Str D Nussloch Germany Phone: Fax: Internet: Assembly contracted to Leica Microsystems Ltd. Shanghai Leica RM2235 1

4 Table of Contents 1. Important Information Safety Safety notes Warnings Integrated safety devices Instrument Components and Specifications Overview instrument components Instrument specifications Technical Data Instrument Setup Standard delivery Installation site requirements Unpacking and installation Assembling the handwheel Inserting the universal cassette clamp Inserting the knife holder Operation Operating elements and their functions Section thickness setting Coarse driving wheel Mechanical trimming function Specimen retraction Specimen holder with precision orientation Adjusting the clearance angle Fine adjustment of the force balance Clamping the specimen Clamping the knife / disposable blade Sectioning Changing the specimen or interrupting sectioning Finishing the daily routine Optional Accessories Assembly for fixture for specimen clamps Rigid fixture for specimen clamps Directional fixture for specimen clamps Instructions for Use V 2.1 RevF 11/2015

5 Table of Contents Fine-directional fixture for specimen clamps Quick clamping system Specimen clamps and holders Standard specimen clamp Vee insert Foil clamp type Universal cassette clamp Super mega-cassette clamp Holder for round specimens Knife holder base and knife holder Knife holder base, without lateral displacement feature Knife holder E/E-TC Knife holder N/NZ Section waste tray Backlighting Tray Freezer pack Universal microscope carrier Magnifying lens Additional accessories Ordering information Troubleshooting Possible faults Instrument malfunctions Troubleshooting Cleaning and Maintenance Cleaning the instrument Maintenance instructions Lubricating the instrument Warranty and Service Decontamination Certificate (Master) Leica RM2235 3

6 1. Important Information Symbols in the text and their meanings Warnings appear in a gray box and are marked by a warning triangle. The package must be kept in a dry environment. Indicates the correct upright position of the package. (5) Notes, i.e. important user information appears in a gray box and is marked by an information symbol. Numbers in parentheses refer to item numbers in illustrations. Manufacturer Date of Manufacture This product fulfills the requirements of the Council's Directive 98/79/EC concerning in vitro diagnostics (IVD) medical devices. In vitro diagnostics (IVD) medical device Observe the Instructions for Use Order No. Serial number The package contents are fragile and must be handled with care. It allows maximum 3 stacks layers. Indicates the temperature range permitted for transporting the package. Minimum -29 C, Maximum +50 C Indicates the temperature range permitted for storing the package. Minimum +5 C, Maximum +50 C Indicates the humidity range permitted for storing and transporting the package. Minimum 10 % r.h., Maximum 85 % r.h Tip-n-Tell indicator to monitor whether the shipment has been transported and stored in upright position according to your requirements. With a pitch of 60 or more, the blue quartz sand flows into the arrow-shaped indicator window and sticks there permanently. Improper handling of the shipment is immediately detectable and can be proven definitively. 4 Instructions for Use V 2.1 RevF 11/2015

7 1. Important Information In the Shockwatch system, a precision glass tube shows shocks or impacts that are above a specified intensity through red coloration. Exceeding a defined acceleration (g value) destroys the surface tension of the liquid in the interior of the tube. This causes the indicator tube to change color. Instrument type All information provided in these Instructions for Use applies only to the instrument type indicated on the title page. A nameplate indicating the instrument serial number is attached at the left side of the instrument. Fig. 1 is provided as an example only and shows a valid nameplate for this instrument. Indicates the item can be recycled where correct facilities exist. Qualification of personnel The Leica RM2235 may be operated by trained laboratory personnel only. All laboratory personnel designated to operate the Leica instrument must read these Instructions for Use carefully and must be familiar with all technical features of the instrument before attempting to operate it. Fig. 1 Designated use The Leica RM2235 is a manually operated rotation microtome for creating thin sections of specimens of varying hardness for use in routine and research laboratories in the fields of biology, medicine and industry. It is designed for sectioning soft paraffin specimens as well as harder specimens, as long as they are suitable for being cut manually. Any other use of the instrument will be considered as improper use! Leica RM2235 5

8 2. Safety The safety and caution notes in this chapter must be observed at all times. Be sure to read these notes even if you are already familiar with the operation and use of other Leica products. 2.1 Safety notes These Instructions for Use include important information related to the operating safety and maintenance of the instrument. The Instructions for Use are an important part of the product, and must be read carefully prior to startup and use and must always be kept near the instrument. These Instructions for Use must be appropriately supplemented as required by the existing regulations on accident prevention and environmental safety in the operator s country. This instrument is built and inspected according to the Safety requirements for laboratory instruments. The current EC Declarations of Conformity can be found on the Internet: To maintain this condition and ensure safe operation, the user must observe all notes and warnings contained in these Instructions for Use. 2.2 Warnings The protective devices located on the instrument and the accessories must not be removed or modified. Only service personnel qualified by Leica may repair the instrument and access the instrument's internal components. The safety devices installed in this instrument by the manufacturer only constitute the basis for accident prevention. Operating the instrument safely is, above all, the responsibility of the owner, as well as the designated personnel who operate, service or clean the instrument. To ensure trouble-free operation of the instrument, make sure to comply with the following instructions and warnings. Warnings Safety notes on the instrument itself Safety notes on the instrument itself, which are marked with a warning triangle, indicate that the correct operating instructions (as defined in these Instructions for Use) must be followed when operating or replacing the item marked. Failure to adhere to these instructions may result in an accident, personal injury, damage to the instrument or accessory equipment. 6 Instructions for Use V 2.1 RevF 11/2015

9 2. Safety Warnings Transport and installation Once unpacked, the instrument may be transported only in an upright position. Do not transport the instrument by holding it by the handwheel grips, coarse driving wheel or the knob for setting the section thickness. The protective devices located on the instrument and the accessories must not be removed or modified. Safety instructions working with the instrument Take care when handling microtome knives and disposable blades. The cutting edge is extremely sharp and can cause serious injuries! Always remove the knife / blade before detaching the knife holder from the instrument. Always put the knives back into the knife case when not in use! Never place a knife anywhere with the cutting edge facing upwards and never try to catch a falling knife! Always clamp the specimen block BEFORE clamping the knife. Safety instructions working with the instrument Prior to manipulating the knife and specimen, or changing the specimen or knife, and during breaks, always lock the handwheel and cover the cutting edge with the knife guard. ALWAYS turn the handwheel clockwise; otherwise, the brake will not work properly. Always wear safety goggles when sectioning brittle specimens. Specimens may splinter! Specimen blocks must NOT be oriented during the retraction phase. If a block is oriented during retraction, the block will advance by the retraction value PLUS the selected section thickness before the next section. This may cause damage to both specimen and knife! Prior to sectioning, check that the specimen is securely clamped in the specimen clamp failure to observe this poses the risk of damaging the specimen. Hazards servicing and cleaning Always lock the handwheel before cleaning! Do not use any solvents containing acetone or xylene for cleaning! Ensure that liquids do not enter the interior of the instrument during cleaning! When using cleaners, please comply with the safety instructions of the manufacturer and the laboratory safety regulations! Leica RM2235 7

10 2. Safety 2.3 Integrated safety devices 5 Locking the handwheel There are two ways of locking the handwheel (12): Using the lever (3) on the right side of the microtome base plate, the handwheel can be braked in almost any position. To brake, rotate the lever in a counterclockwise direction to position. 12 Caution! The braking lever (3) must be exactly in position, so that the handwheel brake is applied correctly. If the lever is moved beyond this point, it is possible that the handwheel is no longer braked. To unlock the handwheel brake turn the lever (3) back to its original position. Position. 3 To lock the handwheel, press the lever (5) outwards and continue to turn the handwheel slowly until it locks exactly in the 12 o'clock position. When using both brake systems at the same time, always move the lever (3) to position first. Otherwise, it may be not be possible to release the lever (5). Fig. 2 8 Instructions for Use V 2.1 RevF 11/2015

11 2. Safety 7 Knife holder N 8 7 Knife guard on the knife holder Each knife holder is equipped with a tightly mounted knife guard (8, 9). This makes it possible to cover completely the cutting edge in every knife or blade position. Knife holder N/NZ Fig. 3 The knife guard (8) of the knife holder N/NZ can be easily positioned via the two handles (7) (Fig. 3). To cover the knife edge, push both cover strips of the knife guard to the center. 11 Knife holder E 9 10 Knife holder E/E-TC The knife guard on knife holder E/E-TC consists of a red foldaway handle (9). To cover the cutting edge, fold the knife guard handle (9) upwards as illustrated in Fig. 4. Knife holder E-TC The clamping levers on the knife holder E are not interchangeable. The two clamping levers (10, 11) must remain in the position shown at all times, as otherwise isolated malfunctions of the knife holder can occur. Clamping lever for the blade (10) at the right, clamping lever for the lateral displacement (11) at the left. 9 Fig. 4 Leica RM2235 9

12 3. Instrument Components and Specifications 3.1 Overview instrument components Tray Universal cassette clamp Handwheel locking mechanism Knife holder base, non-orientable Removable section waste tray Smoothturning Handwheel Lever for activating the Handwheel brake Clamping lever of the knife holder base Directional specimen holder fixture Coarse feed wheel Window for displaying the section thickness Adjusting knob for setting the section thickness Lever for activating the mechanical trimming function Knife holder E Clamping lever for lateral displacement Fig Instructions for Use V 2.1 RevF 11/2015

13 3. Instrument Components and Specifications 3.2 Instrument specifications Basic instrument with mechanical trim function, lateral coarse drive The Leica RM2235 rotary microtome is equipped with a low-maintenance, slack-free micrometer drive, with vertical and horizontal specimen feed realized via low-maintenance cross roller bearings. The instrument is equipped with two independent handwheel locking systems for even greater operating safety. Leica's patented, user-adjustable force balance system compensates centrifugal forces arising while cutting via a pretensioned spring for extremely light handwheel action. Advantage: a heavy counterweight in the handwheel is no longer needed. The spring tension is individually adjustable, corresponding to the weight of the respective attached specimen clamp or specimen. The patented specimen retraction system can be switched on and off by the user. The instrument thus offers all of the advantages of specimen retraction while supporting work in "rocking mode", i.e. without a full handwheel rotation. We recommend disabling specimen retraction when working in "rocking mode". The coarse drive wheel is ergonomically positioned. (For more information on the rotating direction, see chapter 5.1.2) Leica RM

14 3. Instrument components and specifications 3.3 Technical Data General Approvals: The instrument-specific marks are located on the rear panel of the instrument next to the nameplate. Operating temperature range: +10 C to +35 C Temperature range during storage: +5 C to +55 C Relative humidity: max. 80 % non-condensing Storage humidity: < 80 % Section thickness range: µm Section thickness settings: from µm in 1.0 µm increments from µm in 2.0 µm increments from µm in 5.0 µm increments Specimen feed: approx. 24 mm, ± 2 mm Vertical stroke: 70 mm Max. cutting range without retraction: 69 mm (without specimen orientation 1 µm) Maximum sectioning area with retraction: 62 mm The specimen retraction can be turned off manually: approx. 40 µm Dimensions and weight Width (including handwheel): Depth (including waste tray): Height (total): Working height (knife blade): Weight (without accessories): 413 mm 618 mm 305 mm (with tray on the hood) 168 mm (measured from the table) approx. 37 kg Optional equipment and optional accessories Specimen orientation (option) Horizontal: 8 Vertical: 8 Angle of rotation: ± 90 Trimming stages: 10 µm, 30 µm Repositioning of knife holder base North-south: ± 25 mm 12 Instructions for Use V 2.1 RevF 11/2015

15 4. Instrument Setup 4.1 Standard delivery The Leica RM2235 standard delivery includes: 1 Leica RM2235 basic instrument 1 handwheel, complete section waste tray tool set, consisting of: Allen key with handle No Allen key with handle No Allen key No screwdriver 3x50, 186 long bottle (50 ml) of oil for drives, type brush w/magnet dust protective cover Instructions for Use, printed (German/English, with Language CD ) The accessories ordered are included in a separate box. Carefully check the delivery against the packing list and the delivery note. Should you find any discrepancies, please contact your Leica sales office without delay. 4.2 Installation site requirements Stable, vibration-free laboratory table with horizontal, flat table top, as far as possible vibration-free ground. No other instruments nearby which might cause vibrations. Room temperature consistently between + 10 C and + 35 C. Obstruction-free access to the handwheel. Never operate the instrument in rooms with an explosion hazard. Leica RM

16 4. Instrument Setup 4.3 Unpacking and installation Fig. 6 When the instrument is delivered, check the tilt indicators on the packaging. If the arrowhead is blue, the shipment was transported laying flat, was tilted at too great an angle or fell over during transport. Note this on the shipping documents and check the shipment for possible damage. 1 2 Remove the packing strap and the adhesive tape (1). Remove the carton lid (2). Take the accessory carton (optional accessories) (3) and the cartons (4) from the standard scope of delivery. 1 The transport carton and included retaining elements should be kept in case a return shipment is necessary later Fig Instructions for Use V 2.1 RevF 11/2015

17 4. Instrument Setup 4.3 Unpacking and installation (cont.) Take out the fixing module (5). To do so, hold it by the top edge of the module and in the recessed grip (6) and pull it out by pulling upwards Remove the outer carton wall (7). Lift the instrument* (8) by holding it by the base plate at the front and under the instrument on the back and lift it out of the formed cushion (9). 8 9 Never hold the instrument for transport by the handwheel or the rotary knob for section thickness adjustment. (*= the instrument illustration is provided as an example only.) Place the instrument on a stable laboratory table. The two sliding elements (10) located on the rear of the base plate make it easier to move the instrument on the table. To move the instrument, hold it by the front of the base plate, lift it up gently and slide it on its slides. Fig Observe the correct resting angle to the table to avoid pinching your fingers. Leica RM

18 4. Instrument Setup 4.4 Assembling the handwheel The handwheel has to be assembled before attempting to use the instrument. The necessary parts and tools can be found in the toolkit supplied in the delivery. The feather key (4) is loosely placed in the handwheel shaft (1) and fixed in place with a cable tie during transport a Remove the cable tie (3). Caution! Make sure not to lose the feather key! Place the handwheel (2) on the handwheel shaft (1) as shown. Tighten the screw (2a) located in the center hole of the handwheel with a Allen key No. 4 (5). Remove the cover foil from the self-adhesive cover disk and fix the cover disk on the handwheel. Fig Instructions for Use V 2.1 RevF 11/2015

19 4. Instrument Setup 4.5 Inserting the universal cassette clamp 60 There are two versions of the specimen holder fixture, one with and one without specimen orientation, which are interchangeable. The specimen orientation allows for simple position correction of the specimen surface when the specimen is clamped into place You can use the quick clamping system (64) to hold all available accessory specimen clamps (for more information, see Chapter 6 "Optional accessories"). To do so, proceed as follows: Move the specimen head (60) to the upper end position by turning the handwheel (12) and engage the handwheel lock. To release the clamping system, turn the screw (61) of the quick clamping system (64) counterclockwise using an Allen key No. 4 (71). Push the guide (63) of the universal cassette clamp (62) from the left into the quick clamping system (64) as far as it will go. To clamp the cassette clamp turn the screw (61) clockwise as far as it will go. Fig. 10 Since all stage clamps available as accessories are equipped with the same kind of guide on the back, they are inserted in the same way described here using the example of the cassette clamp. Leica RM

20 4. Instrument Setup 4.6 Inserting the knife holder Setting up the knife holder base Release the clamping lever (50) by rotating it counterclockwise. Insert the knife holder base (51) using the notch (52) on the bottom into the T-piece (55) of the microtome base plate (53). To secure the knife holder base, turn the clamping lever (50) clockwise. 52 Enlarged detail: Scale for better repositioning of 51 the knife holder for varying specimen heights The knife holder base (51) can be moved back and forth on the microtome base plate. This allows bringing the knife holder to optimal sectioning position in relation to the specimen. There is a scale (54) on the right side of the microtome base plate. This enables faster and better positioning of the knife holder at the specimen if various combinations of standard specimens and specimen holders are used. The rear edge of the knife holder base (51) functions as the scale reference. Fig Inserting the knife holder Loosen the screw (58) using an Allen key No. 4 (71) until the knife holder (57) can be moved. Place the knife holder (57) with the underside groove onto the T-piece (56) of the knife holder base (51). To clamp, retighten the screw (58). 58 Fig Instructions for Use V 2.1 RevF 11/2015

21 5. Operation 5.1 Operating elements and their functions Section thickness setting The section thickness is set by turning the adjusting knob (33) at the front of the microtome on the right. The adjusting knob has a notch for each value that can be set. Setting range: 1-60 µm from 1-10 µm in 1 µm increments from µm in 2 µm increments from µm in 5 µm increments. The section thickness set in each case is displayed in the window (34). The selected section thickness (on the scale) must agree with the red pointer (38) Coarse driving wheel Fig. 13 The instrument can be ordered with clockwise or counterclockwise rotation. The given direction of rotation means "forwards" and relates to the feed movement of the specimen towards the knife. The coarse motion serves for a fast horizontal forwards movement of the specimen - towards the knife - and backwards - away from the knife. When reaching the rear/front end positions, the coarse driving wheel can only be turned with difficulty. In the front end position, no more feed motion takes place. Fig. 14 The coarse driving wheel also turns during sectioning. Therefore it must not block whilst the handwheel is being turned during sectioning; otherwise, no feed motion can take place and thus also no sectioning. Leica RM

22 5. Operation Magnet for holding the wrench size 4 For turning the retraction Off, turn the slot into the horizontal position (off). For turning the retraction On, turn the slot into the vertical position (on). Fig Specimen retraction The specimen retraction serves for protecting the knife and the specimen. When the retraction is switched on, the specimen is drawn back 40 µm into the upper end position after the sectioning stroke during the return movement. Before the feed motion of the new section thickness, the feed motion for the retraction value takes place. The specimen retraction can be switched off manually at the back of the instrument (Fig. 15), if required, using the slotted-screwdriver supplied with the delivery. Before switching the specimen retraction On and Off, run the specimen head to the upper end position by turning the handwheel Mechanical trimming function 36 Fig. 16 The RM2235 is fitted with a mechanical trimming function. The trimming lever has 3 notching positions (0, 10 µm, 30 µm). The points (36) mark the two trimming stages: = 10 µm = 30 µm For activating the trimming function, press the lever downwards into one of the two notching positions and keep depressed. After each rotation of the handwheel, a feed motion of 10 µm or 30 µm takes place. After letting go of the lever, it automatically springs back to its original position (zero position). The trimming function is thereby deactivated. The section thickness that has been set is not added to the selected trimming value. If the section thickness that has been set is greater than the selected trimming value, the section thickness is fed. 20 Instructions for Use V 2.1 RevF 11/2015

23 5. Operation Specimen holder with precision orientation In the quick clamping device of the specimen holder fixture with precision orientation, all specimen clamps available as optional accessories can be used (implemented) The specimen orientation allows for simple position correction of the specimen surface when the specimen is clamped into place. Orienting the specimen Specimen blocks must NOT be oriented during the retraction phase! If a block is oriented during retraction, the block will advance by the retraction value PLUS the selected section thickness before the next section. This may cause damage to both specimen and knife! Display of the zero position Fig. 17 For better display of the zero position, the orientation has two red indicators (32). When both indicators are visible and both setscrews are in zero position at the same time (notch point, white marking on " "), the specimen is in zero position. When the large standard specimen clamp (50 x 55 mm) is used, the specimen orientation of 8 in north-south direction is no longer possible. The usable angle is only about 4 in this case. Raise the specimen head to the upper end position and activate the handwheel lock. To release the clamp, turn the eccentric lever (29) forwards. Turn setscrew (30) to orient the specimen in north-south direction. Turn setscrew (31) to orient the specimen in east-west direction. Each complete turn of the screw inclines the specimen by 2. A total of 4 complete turns = 8 are possible in every direction. The accuracy is approximately ± 0.5. For ease of estimation, there is a white mark on the handle and a click stop that is noticeable during turning. To lock the current orientation, turn the eccentric lever (29) backwards. Leica RM

24 5. Operation 5.2 Adjusting the clearance angle The index marks (0, 5 and 10 ) for adjustment of the clearance angle (59.1) are located on the right side of the knife holder (57) There is also an index mark (59.2) on the right side of the knife holder base (51) which serves as a reference point when adjusting the clearance angle. 58 Loosen the screw (58) using an Allen key No. 4 (71) until the knife holder (57) can be moved. Enlarged detail: Index marks for adjusting the clearance angle. Move the knife holder until the index mark of the desired clearance angle coincides with the reference line on the knife holder base. Example: Enlarged detail showing a clearance angle setting of The recommended clearance angle setting for knife holder E is approx. 5. The usable angle is only about 4 in this case. Hold down the knife holder in this position and retighten the screw (58) for clamping. Fig Instructions for Use V 2.1 RevF 11/2015

25 33 Important! Never turn the screw more than 1 / 2 turn at a time. Fig Operation 5.3 Fine adjustment of the force balance If another accessory of a different weight is mounted on the specimen head (33), you must check whether it is necessary to readjust the force balance. Checking the correct setting: Attach the new accessory and clamp the specimen. Set the specimen head to half the height of the vertical travel range by turning the handwheel (Fig. 19). If the specimen head remains in this exact position, the setting is correct. If the specimen head moves, i.e. it is raised or lowered, fine adjustment is necessary. Failure to adjust the force balance may result in injury while working. The force balance is adjusted using the screw (34), which can be accessed by removing the section waste tray on the bottom of the base plate of the microtome. Use the Allen key provided, No. 5 (with handle) for the adjustment. If the specimen head moves downwards, turn the screw approx. 1 / 2 turn clockwise. If the specimen head moves upwards, turn the screw (34) approx. 1 / 2 turn counterclockwise. Continue this procedure until the specimen head no longer moves once released. 34 Fig. 20 Leica RM

26 5. Operation 5.4 Clamping the specimen Always clamp the specimen block BEFORE clamping the knife or the blade. Lock the handwheel and cover the knife edge with the knife guard prior to any manipulation of knife or specimen, prior to changing the specimen block and during all work breaks! Rotate the handwheel until the specimen clamp is in the uppermost position. Block the handwheel (allow lever (5) Fig. 2 to notch) and activate the brake. Insert a specimen into the specimen clamp. A detailed description for inserting the specimen into various specimen clamps and specimen holders is provided in Chapter 6 "Optional accessories". 5.5 Clamping the knife / disposable blade Take care when handling microtome knives and disposable blades. The cutting edge is extremely sharp and can cause serious injuries! Carefully insert knife or disposable blade into the knife holder and clamp. Make sure that the blade is clamped parallel to the upper edge of the pressure plate. (For more information see Chapter 6.3.2, Fig. 33) A detailed description for inserting the blade or the knife into the individual knife holders is provided in Chapter 6, "Optional accessories". 24 Instructions for Use V 2.1 RevF 11/2015

27 5. Operation 5.6 Sectioning Cutting into the specimen (trimming) Always turn the handwheel evenly in clockwise direction; otherwise, the brake will not work properly. The rotation speed of the handwheel must be adapted to suit the hardness of the specimen. For harder specimens, use a slower speed. Remove the sections Run the specimen to the rear end position by turning the coarse driving wheel. Push the knife holder on the knife-holder base almost until it is just before the specimen. Orientate the position of the specimen surface (only in the case of specimen holders that can be orientated). Release the handwheel lock, or handwheel brake, respectively. Using the trimming lever select the required trimming stage. Begin the cutting process by turning the handwheel. Stop the cutting process when the required specimen level has been reached. Let go of the trimming lever. Set the required section thickness, or check the value setting on the display, respectively. Always use a different area of the cutting edge for trimming and sectioning. To do so, laterally displace the blade or knife in the knife holder. When using the knife holder E with lateral displacement, it is sufficient to move the knife holder sideways. For sectioning, turn the handwheel evenly in a clockwise direction. Take care not to block the coarse driving wheel when turning the handwheel! Otherwise there will be no feed motion of the section thickness and thus no sectioning will occur. Pick up the sections and mount them on microscope slides. Leica RM

28 5. Operation 5.7 Changing the specimen or interrupting sectioning Lock the handwheel and cover the knife edge with the knife guard prior to any manipulation of knife or specimen head, as well as prior to changing the specimen block and during all work breaks! Raise the specimen to the upper end position and activate the mechanical handwheel lock. Cover the sectioning edge with the knife guard. Remove the specimen from the specimen clamp and mount a new sample to continue. Run the specimen clamps with the coarse driving wheel back far enough until the new specimen can start being cut. 5.8 Finishing the daily routine Move the specimen to the upper end position by turning the handwheel and engage the handwheel lock. Always remove the knife / blade before detaching the knife holder from the instrument. Always put the knives back into the knife case when not in use! Never place a knife anywhere with the cutting edge facing upwards and never try to catch a falling knife! Remove the blade from the knife holder and insert it in the receptacle at the bottom of the dispenser, or remove the knife from the knife holder and put it back in the knife case. Remove the specimen from the specimen clamp. Push all section debris into the section waste tray and empty the tray. Clean the instrument (see Chapter 8.1). 26 Instructions for Use V 2.1 RevF 11/2015

29 6. Optional Accessories 6.1 Assembly for fixture for specimen clamps Depending upon the purchase order, the basic instrument is delivered with the directional or rigid fixture for specimen clamps which must be assembled first. All specimen clamps available as accessories can be used in both fixtures for specimen clamps. Before assembling the fixture for specimen clamps, activate the mechanical handwheel lock! Rigid fixture for specimen clamps Screw the rigid fixture for specimen clamps (4) onto the specimen head (3): Remove the screw (1), place the specimen holder fixture (4) onto the specimen head (3) from the front and tighten the screws (2) with an Allen key No. 3. Next, insert the screw (1) from the side and briefly tighten it with an Allen key No Fig. 21 Remove the rubber ring only after attaching the specimen head! 1 2 Fig. 22 Leica RM a 5b b 9b 9a 9a Finally, place the dovetail holder (2) and fasten by screwing in the 4 screws (1) using an Allen key No Directional fixture for specimen clamps Loosen the eccentric bolt (6) by turning it counterclockwise. Completely unscrew the thrust piece (5) with a flat-tip screwdriver and pull it out with spring (5a) and pin (5b). Completely unscrew the setscrews (3) and (4). Attach the directional fixture for specimen clamps as shown. Insert the screws (7+8) in the bore (2 screws (8) are accessible through the bore (9a) and evenly screw them in using an Allen key No. 3. Insert the spring (5a) and pin (5b) with the flatter side into the thrust piece (5). Completely screw in the thrust piece with a flat-tip screwdriver. Completely screw in the setscrews (3+4). 27

30 6. Optional Accessories Fine-directional fixture for specimen clamps Before the fine-directional fixture for specimen clamps can be mounted, loosen 4 screws (10) (Allen key No. 3) and carefully remove the fixture for specimen clamps from the baseplate (9) Using the 4 supplied screws (11) and the Allen key No. 3, fasten the baseplate to the specimen head (12). 11 Now, screw the fine-directional fixture for specimen clamps with the 4 screws (10) and the Allen key No. 3onto the specimen head. If the fine-directional fixture for specimen clamps is not used, retain the baseplate and 4 screws (11) together with the fine-directional fixture for specimen clamps! Fig Instructions for Use V 2.1 RevF 11/2015

31 6.1.4 Quick clamping system 6. Optional Accessories It is used as specimen holder for use with the fine-directional fixture for specimen clamps with zero point indicators or the directional fixture for specimen clamps. Screw the 4 screws (13) into bore A with an Allen key No. 2.5 and tighten them. a 13 a Fig. 24 Leica RM

32 6. Optional Accessories 6.2 Specimen clamps and holders All specimen clamps available as accessories can be integrated into either the directional or non-directional specimen holder fixture Standard specimen clamp The standard specimen clamp is available in two sizes: 40 x 40 mm and 50 x 55 mm. They are designed for direct clamping of rectangular blocks. In addition, they accommodate the foil clamps. Turn the knurled screw (66) counterclockwise to move the movable jaw (68) downward. Mount the specimen (67) as required. Turn the knurled screw (66) clockwise to move the movable jaw upward against the fixed jaw to securely clamp the sample. Fig. 25 When clamping cassettes, make sure that they are not clamped too tightly, since the cassette bodies can bend and result in sections that are too thick or thin, or the entire specimen may fall out and become damaged. 30 Instructions for Use V 2.1 RevF 11/2015

33 6. Optional Accessories Vee insert The vee insert (70) is mounted in the hole provided in the lower movable jaw of the standard specimen clamp. This makes it possible to clamp round specimens in the standard specimen clamp. Turn the knurled screw (66) counterclockwise to move the movable jaw (68) downward. Insert the pin (70.1) of the vee insert (70) in the hole (69) of the lower jaw (68). Mount the specimen as required. Turn the knurled screw (66) clockwise to move the movable jaw with the vee insert upward against the fixed jaw to securely clamp the sample. Fig. 26 Leica RM

34 6. Optional Accessories Foil clamp type 1 The foil clamp type 1 is appropriate both for clamping very small and thin foil pieces and flat, angular specimens. It is mounted in the standard specimen clamp Clamping of foil pieces Move the movable jaw (74) to the right as required by turning the setscrew with an Allen key No. 4 (71). Place the foil (72) between the movable jaw (74) and the fixed jaw (73). To clamp the foil, screw the movable jaw (74) against the fixed jaw (73) by using the Allen key. Insert the foil clamp (75) in the standard specimen clamp as shown. Turn the knurled screw (66) clockwise to clamp the foil clamp in the standard specimen clamp Clamping of flat, angular specimens To clamp angular specimens, replace the long setscrew (76) with the short setscrew (77) provided with the foil clamp. Fig. 27 Unscrew the long setscrew (76) to the left with an Allen key size 4 (71). Screw the short setscrew (77) in the hole. Place the sample (67) between the movable jaw (74) and the fixed jaw (73) To clamp the sample, screw the movable jaw (74) by turning the setscrew (77) against the fixed jaw (73). Insert the foil clamp in the standard specimen clamp as shown. 77 Turn the knurled screw (66) clockwise to clamp the foil clamp in the standard specimen clamp. 67 Fig Instructions for Use V 2.1 RevF 11/2015

35 6. Optional Accessories Universal cassette clamp 60 Prior to sectioning, laboratory personnel MUST check that the cassette is seated securely in the universal cassette clamp. 65 Push the lever (60) forwards. Mount the cassette (65) horizontally or vertically as required. To clamp the cassette, release the lever (60). Fig. 29 Leica / Surgipath cassettes with minimum dimensions of 39.8 x 28 mm and maximum dimensions of 40.9 x 28 mm may be clamped in the universal cassette clamp (UCC) horizontally as well as vertically. When using other particularly thin-walled cassettes, the cassette may become deformed or other problems may arise with the clamping system. If the user tries clamping the cassette and realizes that it is not securely clamped into place, an alternative tensioning clamp must be used. When using cassettes whose lid is molded on, make sure that the broken edge left by removing the lid does not prevent the specimen from being securely clamped if necessary, the specimen must be clamped horizontally. Before clamping the cassette into the universal cassette clamp, remove excess wax on the outside of the cassette to ensure that the cassette clamps in securely. Wax deposits on the outside of the cassette can make the universal cassette clamp dirty. The dirt prevents the cassette from clamping in securely and can lead to sections being too thick or thin, chattering within the section and, in the worst-case scenario, damage to the specimen. Prior to sectioning, the user has to verify that the specimen is clamped securely and, if necessary, remove wax deposits from the universal cassette clamp according to the specifications in Chapter 8.1 "Cleaning and maintenance Universal cassette clamp". Leica RM

36 6. Optional Accessories Super mega-cassette clamp Fig. 30 Assembly of the super mega-cassette clamp The super mega cassette clamp should preferably be used with the rigid fixture for specimen clamps. To do so, proceed as follows: Remove the rubber ring (5) only after having attached the cassette clamp on the specimen head! Screw the rigid fixture for specimen clamps (4) onto the specimen head (3): Remove the screw (1), place the specimen holder fixture (4) onto the specimen head (3) from the front and tighten the screws (2) with an Allen key No. 3. Next, insert the screw (1) from the side and briefly tighten it with an Allen key No. 4. Insert the super mega cassette clamp from the side on the left into the dovetail guide of the rigid fixture for specimen clamps and tighten screw (1). If the directional fixture for specimen clamps is used with the rigid knife holder base, the orientation must be in position "0" and the cover for backlighting illumination must be detached. (Danger of collision if not observed!) NEVER use the super mega-cassette clamp with backlighting illumination! 34 Instructions for Use V 2.1 RevF 11/2015

37 6. Optional Accessories Holder for round specimens The holder for round specimens is designed to accommodate cylindrical specimens. Inserts for specimens of 6, 15 and 25 mm diameter are available To mount the required insert (89.1-3) turn the clamping ring (90) counterclockwise and remove it Place the required insert into the tension ring (90) and screw the tension ring onto the thread (91) by turning it clockwise. 91 Mount the specimen (67) and clamp it by turning the clamping ring (90) clockwise To orient the inserted specimen, insert the pin (92) into the bore (93) and rotate it counterclockwise to release the clamp. You can now rotate the specimen so that the side you want faces upwards. To lock it in the position you have chosen, tighten the pin (92) by turning it clockwise. Fig. 31 Leica RM

38 6. Optional Accessories 6.3 Knife holder base and knife holder The plastic handles of all clamping levers on the instrument and knife holders can be turned to the position that is most convenient for each user. 94 Pull the grip (94) out of the lever, hold it in this position, and rotate it to the desired position. It will then lock automatically when released. Fig Knife holder base, without lateral displacement feature Repositioning the knife holder base The one-piece knife holder base (rigid) (51) can be moved forwards and backwards on the microtome base plate. This vertical displacement allows bringing the knife holder into the optimal cutting position in relation to the specimen. 51 To release, rotate the clamping lever (50) on the right side of the microtome base plate counterclockwise. Reposition the knife holder together with the knife holder base forward or backward as appropriate. Secure the clamping mechanism by rotating the lever (50) clockwise. 50 Fig Instructions for Use V 2.1 RevF 11/2015

39 6. Optional Accessories Knife holder E/E-TC The knife holder E-TC is designed for the Leica TC-65 tungsten carbide blades. Knife holder E-TC Prior to inserting the blade, both knife holder and knife holder base must have been installed on the instrument! Inserting the blade, knife holder E and E-TC Fold knife guard (9) downward. To insert the blade, flip the right clamping lever (10) forward and down. Carefully insert the blade from the side. Make sure that the blade is clamped parallel to the upper edge of the pressure plate. To clamp the blade, rotate clamping lever (10) back upwards Fig. 34 The knife holder E is designed for conventional disposable blades from all current manufacturers. It is available in two models: one for low-profile blades and one for high-profile blades. The knife holder has a lateral displacement, so that the entire width of the blade can be used. 11 Knife holder E 9 10 The clamping levers on the knife holder are not interchangeable. The two clamping levers (10, 11) must remain in the position shown at all times, as otherwise isolated malfunctions of the knife holder can occur. Clamping lever for the blade (10) at the right, clamping lever for the lateral displacement (11) at the left. Fig. 35 Leica RM

40 6. Optional Accessories Knife holder E 10 Lateral displacement (only for knife holder E) The lateral displacement feature of the knife holder base enables the use of the entire length of the blade, eliminating the need for readjusting the knife holder. The knife holder E consists of a segment arch A (with lever (11)) and the clamp mount B (with lever (10) and ejector (13)). The extreme left and right positions as well as the middle position are each marked with a click stop. a B 12 Fig. 36 To release the clamp, rotate the lever (11) on the left side of the knife holder forwards. Move the knife holder sideways. To clamp, rotate the lever (11) back. Note on the knife holder E: To ensure a proper sectioning result, clamp mount B must be securely clamped onto segment arch A. The clamping is carried out using an eccentric at the lever (11). The clamping force is adjusted with the setscrew (12) on the underside of the segment arch. The setting of the clamping is carried out so that the clamping lever can be rotated to the stop with constantly increasing resistance. The knife holder E is an important precision component, the quality and precise adjustment of which have a lasting effect on the entire function of the microtome. In case of malfunctions or damages to the clamp mount, it is always required to replace the clamp mount, including the pertinent clamping lever. Leica Biosystems offers special prices for new clamp mounts in case of damages to the clamp mount after the warranty has expired. In this way, perfect function of the device can be ensured over the course of many years. Setting of the clamping system of the clamp mount on the segment arch Adjust the clamping with a 2.5 mm Allen key at the setscrew (12) so that the lever initially "slips" when the lever is activated. Continue turning the setscrew (12) a little bit (approx. 1 / 4 turn to the left or right) then check that the lever no longer "slips", but also does not jam "heavily". (The language CD features a short video film for this purpose.) 38 Instructions for Use V 2.1 RevF 11/2015

41 6. Optional Accessories Knife holder E with a water trough for low-profile blades The knife holder E with a water trough is only for low-profile blades. The knife guard on knife holder E consists of a red foldaway handle (9). To cover the cutting edge, fold the knife guard handle (9) upwards as illustrated in the figure. Fig. 37 The clamping levers on the knife holder are not interchangeable. The two clamping levers (10 and 11) must remain in the position shown at all times, as otherwise isolated malfunctions of the knife holder can occur. Clamping lever for the blade (10) at the right, clamping lever for the lateral displacement (11) at the left. Use Floating thin paraffin sections (for example, for subsequent immunostaining procedures) on the surface of the water. Flat paraffin sections can be removed from the surface of the water using glass slides. The vessel is filled with water up to the blade. After trimming, remove the section waste from the tray and create the sections to be prepared. The sections floating on the surface of the water can be removed using the glass slide. Fig. 38 Leica RM

42 6. Optional Accessories Knife holder N/NZ The knife holders N and NZ are appropriate for standard steel and tungsten carbide knives, profile c and d, up to 16 cm long. The integrated height adjustment feature allows you to also use knives that have been resharpened numerous times. Mounting the knife support bar Push knife guard (8) to the center. Set the knife support bar (46) onto the height adjustment screws (not visible) in the position shown. The flat ends of the height adjustment screws must be located in the slots at each end of the knife support bar. Prior to inserting the knife, both knife holder and knife holder base must have been installed on the instrument! Knife holder N For holding conventional knives up to 16 cm in length. Enlarged detail: Knife inserted and height-adjusted Knife holder NZ For holding conventional and carbide metal knives up to 16 cm in length. Knife pressure plate (56) for extreme stability and full utilization of the knife blade Fig Instructions for Use V 2.1 RevF 11/2015

43 6. Optional Accessories Inserting the knife Rotate the knurled nuts (48) on the right and left of the knife holder forward in opposite directions, lowering the knife support bar to the lowest possible position, thus ensuring that the knife edge will not be damaged when inserting the knife. Unscrew the clamping screws (49) as far out as possible (rotate counterclockwise). Hold the knife (47) at the knife base and carefully insert it in the holder from the side as shown with the cutting edge facing upward. Knife height adjustment Lateral repositioning of the knife When adjusting the clearance angle, the knife edge should be positioned as exactly as possible in the actual center of rotation of the knife holder. The lay-on edge (39) of the rear clamping chucks serves as a reference position for correct knife height adjustment. The knife edge should be parallel with the locating edges. Rotate the knurled nuts (48) uniformly and backwards until the knife blade is parallel to the lay-on edge (39) (see detailed illustration) of the rear clamping chucks. To clamp the knife (47) evenly screw the two knife clamping screws (49) inward (rotate clockwise). Push knife guard (8) to the center. Loosen the clamping screws (49) by turning them counterclockwise. Push the knife (47) to the left or right as required. To clamp the knife (47), always tighten the clamping screw (49) first which is located on the side to which the knife has been repositioned by turning it clockwise. Leica RM

44 6. Optional Accessories 6.4 Section waste tray 53 Push the section waste tray (18) from the front to the microtome base plate (53) until it is held in place by the two magnets (39) (on the front of the microtome base plate). To remove the section waste tray, lift it slightly and pull it off towards the back Fig Backlighting The backlighting is inserted into the front of the one-piece knife holder base Remove the two screws (1) using a slotted screwdriver and then remove the cover plate (2). Insert the backlighting (3) in the recess at the rear of the knife holder base. Insert the plug (4) for the backlighting into the socket (5) of the microtome and connect the plug of the power adapter to an AC power socket. Fig. 41 The backlighting illuminates once the microtome is turned on with the power switch. NEVER use the backlighting illumination with the super mega-cassette clamp! 42 Instructions for Use V 2.1 RevF 11/2015

45 6. Optional Accessories 6.6 Tray The tray is mounted on the hood of the microtome so that the small feet on the underside fit into the cutouts on the hood. It is for storage of the utensils used during sectioning as well as the sectioned specimens. Fig Freezer pack The freezer pack consists of the freezer plate (98) and the insulation jacket (99). It is for cooling (or keeping cold) paraffin specimens. Up to 35 standard cassettes can be cooled at the same time. The specimens require approx. 20 min. for being cooled from room temperature (approx. 20 C) to "sectioning temperature". The cooling performance depends on the ambient temperature and the volume of the specimens being cooled. The insulation jacket can be placed on the hood of the microtome in the same manner as the tray, or it can be placed next to the instrument. When using the freezer pack on the instrument the tray (Fig. 42) must be removed. 99 Fig. 43 Remove the freezer plate from the insulation jacket and place it in a deep freezer, ideally overnight (but for at least six hours) at approx. -23 C. Reassemble the freezer plate and insulation jacket and place them on the bench or the instrument. Leica RM

46 6. Optional Accessories 6.8 Universal microscope carrier a Unpack all accessories in the package and check for completeness d e 4 Fig Baseplate with drilling (a) 2 - Vertical column with size 8 cap screw (b) and lock washer (c) 3 - Horizontal arm with cross-member (d) and support ring (e) 4 - Support plate, large (for RM2235, RM2245 and RM2255) 5 - Support plate, small (for RM2265) 6 - Allen key size countersunk screws to install support plate 8 - Allen key size 8 c b 5 Fig. 45 Assembling the universal microscope carrier 1. Attach baseplate Select the large (5) or small (4) support plate, depending on the microtome to be used. Attach the support plate to the baseplate with the included 4 countersunk screws (7) using the Allen key No. 3 (6). Fig Instructions for Use V 2.1 RevF 11/2015

47 6. Optional Accessories c b Fig Attach vertical column Insert the cap screw (b) into the hole of the baseplate from below. Place the lock washer (c) on the cap screw from above. Thread the silver vertical column (2) onto the baseplate from above and tighten with the Allen key No The lock washer must be positioned between the baseplate and vertical column to prevent unintentional turning of the column. a Extreme danger of tipping! After installing the vertical column, immediately place the microtome onto the baseplate so that the front feet of the microtome are positioned in the shallow recesses (A). f Fig. 48 h 2 d e g 3. Attach horizontal arm Slide the support ring (e) onto the vertical column and position it so that the lock nut (f) is facing to the back. Tighten the lock nut. Slide the cross-member (d) onto the column. Ensure that the lock screw (g) is facing to the right of the baseplate. The horizontal arm must be centered over the microscope. Slide the horizontal arm (3), flat side facing the lock screw (g), into the cross-member (d) and tighten. Fig. 49 For more information on connecting and using the microscope, magnifying lens or cold-light source, please see the appropriate Instructions for Use. Leica RM

48 6. Optional Accessories 6.9 Magnifying lens 1 2 The magnifying lens provides a 2x magnification and can be used with all Leica 2200 series rotary microtomes. 4 Fig. 50 Open the screw (3) on the horizontal arm of the microscope carrier in a counterclockwise direction. Insert the silver connection piece (1) as far as it will go. Tighten the screw (3). The adapter (2) allows a fiber-optic light guide to be installed. 3 The instrument illustrations in Fig. 51 and Fig. 52 are an example only. Fig. 51 Adjust the position of the magnifying lens to the specimen being processed. The magnifying lens can be swiveled completely to the side if necessary. Always protect the magnifier glass from exposure to direct sunlight! Exposure to sunlight may cause a burning glass effect. Danger of fire! Fig. 52 Use the provided protective cap (4) to cover the magnifying lens. 46 Instructions for Use V 2.1 RevF 11/2015

49 6. Optional Accessories 6.10 Additional accessories Knife holder base, non-orientable Silver for knife holders N, NZ, E, and E-TC Order No Fig. 53 Knife holder N, silver, for holding conventional knives up to 16 cm in length, height adjustment of the knife blade, separate clearance angle adjustment, movable knife guard. Order No Fig. 54 Knife holder NZ, silver for holding conventional and carbide metal knives up to 16 cm in length, knife pressure plate for extreme stability and full utilization of the knife blade, height adjustment of the knife blade, separate clearance angle adjustment, movable knife guard. Order No Fig. 55 Leica RM

50 6. Optional Accessories Fig. 56 Knife holder E, for low-profile disposable blades, (80 x 8 x 0.25 mm), silver, for RM22xx Quick clamp system with ability to shift the blade laterally. 3 click-stop positions make it easier to utilize the entire blade. Color-coded, foldaway knife guard. With blade ejector Order No For high-profile disposable blades, (80 x 14 x 0.35 mm), silver, for RM22xx Order No Fig. 57 Knife holder E, for low-profile disposable blades, (80 x 8 x 0.25 mm), silver, for RM22xx with a water trough, Quick clamp system with ability to shift the blade laterally. 3 click-stop positions make it easier to utilize the entire blade. Color-coded, foldaway knife guard. With blade ejector Order No Instructions for Use V 2.1 RevF 11/2015

51 Knife holder E-TC, 6. Optional Accessories for carbide metal disposable blades TC-65, silver Quick clamp system, rustproof clamping plate made of stainless steel, rear pressure plate made of carbide metal Order No Fig. 58 Disposable blades low profile (819) 80 x 8 x 0.25 mm 01 package of 50 pcs packages of 50 pcs Fig. 59 Disposable blades high-profile (818) 80 x 14 x 0.35 mm 01 package of 50 pcs packages of 50 pcs Fig. 60 Leica RM

52 6. Optional Accessories Fig. 61 Disposable blades Leica TC-65 Leica TC-65 microtome, disposable blade system for sectioning hard specimen materials. The Leica TC-65 carbide metal disposable blades were specially developed for the requirements in labs where hard, blunt materials are routinely sectioned. The one-of-a-kind fine-grain carbide metal guarantees sections to approx. 1 µm. The blades are fully recyclable. Length: 65 mm Thickness: 1 mm Height: 11 mm 1 pack of 5 pcs. Order No Knife 16 cm - profile c - steel Knife, 16 cm long, profile c Note: Knife case included Order No Profile Knife 16 cm, profile d - steel Knife, 16 cm long, profile d Note: Knife case included Order No Fig Instructions for Use V 2.1 RevF 11/2015

53 6. Optional Accessories Knife, 16 cm, profile d, tungsten carbide Knife, 16 cm long, tungsten carbide, profile d Note: Knife case included Order No Fig. 63 Knife 16 cm, profile c, tungsten carbide Knife, 16 cm, tungsten carbide, profile c Note: Knife case included Order No Carbide metal knife serial number Fig. 64 Knife case Variable knife case (plastic), for 1 or 2 knives: cm long (Carbide metal or SM2500 knife: only for 1 knife!) Order No Fig. 65 Leica RM

54 6. Optional Accessories Universal cassette clamp with adapter RM2125 RTS, silver For use with standard cassettes with dimensions that are 39.8 x 28 mm at minimum and 40.9 x 28 mm at maximum. Order No Fig. 66 Standard specimen clamp 50 x 55 mm, with adapter, silver Order No Fig. 67 Standard specimen clamp 40 x 40 mm, with adapter, silver Order No Fig Instructions for Use V 2.1 RevF 11/2015

55 6. Optional Accessories specimen holder fixture, directional silver For holding the standard clamp, universal cassette clamp, round specimen holder and EM specimen holder, including installation hardware Note: Quick clamping system has to be ordered separately Order No Fig. 69 Specimen holder fixture, non-directional silver, for holding the standard clamp, universal cassette clamp, round specimen holder and EM specimen holder, including installation hardware Order No Fig. 70 Specimen holder fixture, fine directional, silver, with 2 zero point indicators, XY orientation 8 per direction, click stops every 2 Note: Quick clamping system must be ordered separately. Order No Fig. 71 Leica RM

56 6. Optional Accessories Quick clamping system, for specimen holder for use with the fine-directional fixture for specimen clamps with zero point indicators or the directional fixture for specimen clamps Order No Fig. 72 Round specimen holder for 6, 15 and 25 mm specimen diameters, with adapter, with 3 clamping rings, silver Order No Fig. 73 Vee insert for standard specimen clamp, silver Order No Fig Instructions for Use V 2.1 RevF 11/2015

57 Foil clamp Type I 6. Optional Accessories for standard specimen clamp, black Maximum specimen size: 25 x 13 mm Order No Fig. 75 Special wrench, for EM specimen holder Order No Fig. 76 EM specimen holder fixture, for EM specimen holder, black For samples with a 10 mm diameter Order No Fig. 77 Leica RM

58 6. Optional Accessories EM universal specimen holder, For samples with a 8.5 mm diameter Order No Fig. 78 EM flat specimen holder, Opening width up to 4.5 mm Order No Fig. 79 Super Mega cassette clamp, with adapter, silver Fig. 80 Note: Use only together with non-orientable specimen holder fixture , which has to be ordered separately. Backlighting cannot be used in connection with a Super Mega cassette clamp. We recommend use with Surgipath Super Mega cassettes and covers, white (VSP 59060B-BX, VSP CS) and Super metal embedding molds (VSP58166) (L x W x H) 75 x 52 x 35 mm Order No Instructions for Use V 2.1 RevF 11/2015

59 6. Optional Accessories Leica RM CoolClamp Fig. 81 Electrically cooled universal cassette clamp with adapter for the Leica RM2200 series rotary microtomes. For use with standard cassettes with dimensions that are 39.8 x 28 mm at minimum and 40.9 x 28 mm at maximum. Uniform microtome sections through electric cooling of the universal cassette clamp using the Leica RM CoolClamp. Energy-saving cooling through the heat dissipation system. The patented force balance system of the Leica RM2200 series rotary microtomes enables the specimen clamps to be changed quickly and reliably without modifying the handwheel. Antistatic material makes cleaning easier. Can be retrofitted for all rotary microtomes from the Leica RM2200 series. Technical data: Precooling time until work begins: 30 minutes Temperature: 20 K below ambient temperature Ambient temperature range: +10 C to +35 C Relative humidity: max. 80 %, non-condensing Weight: approx. 650 g Measurements (W x D x H): 80 x 114 x 204 mm Power Inlet: V/ 50/ 60 Hz Certifications: CE, c_csa_us Standard delivery: Please note: The RM CoolClamp cannot be used in connection with a microscope or magnifier. RM CoolClamp Power supply with cable and 3 adapters (UK, US, EU) 5 cable clamps Instructions for Use and DVD Order No Leica RM

60 6. Optional Accessories Magnifier, For assembly on microscope carriers ( ), 2x magnification Note: Magnifier includes an adapter for holding the optional fiber-optic light guide ( ) Order No Fig. 82 Universal microscope carrier, universal assembly Order No Fig. 83 Light guide, gooseneck Order No Fig Instructions for Use V 2.1 RevF 11/2015

61 Leica CLS 100, cold light source, 6. Optional Accessories with power adapter USA V / Hz Order No with power adapter Europe and the United Kingdom V / Hz Order No with power adapter Australia 240V / Hz Order No Fig. 85 Backlighting, Note: For RM2235 and RM2245 only together with external power supply Backlighting cannot be used in connection with a Super Mega cassette clamp Order No Fig. 86 External power supply unit, for use with backlighting , for rotary microtome series Leica RM2235 and RM2245 The following adapters are included: UK, Europe, USA/Japan, Australia Fig. 87 Order No Leica RM

62 6. Optional Accessories Tray, for rotary microtome series Leica RM2200 Order No Fig. 88 Cooling plate, with one freezer pack Order No Fig. 89 Section waste tray, large, included in the standard scope of delivery of RM2235, RM2245, and RM2255. Order No Small, included in the standard scope of delivery of 2265 Order No Fig Instructions for Use V 2.1 RevF 11/2015

63 6. Optional Accessories Brush, "Leica" with magnet for blade removal tool for knife holder E. Order No Fig. 91 Leica RM

64 6. Optional Accessories 6.11 Ordering information Knife holder-base rigid, silver Knife holder N RM22xx, silver Knife holder NZ RM22xx, silver Knife holder E low profile RM22xx, silver, with ejector Knife holder E high profile RM22xx, silver, with ejector Knife holder E with a water trough for low profile blades RM22xx Knife holder E-TC RM22xx silver Disposable blades low-profile, 1 pack of 50 (80 x 8 x 0.25 mm) Disposable blades low-profile, 10 packs of Disposable blades high-profile, 1 pack of 50 (80 x 14 x 0.35 mm) Disposable blades high-profile, 10 packs of Disposable blades Leica TC Knife 16 cm - profile c - steel Knife 16 cm - profile d - steel Knife 16 cm - profile d - carbide metal Knife 16 cm - profile c - carbide metal Universal cassette clamp, with adapter RM2125 (silver) Standard specimen clamp with adapter 50 x 60 (silver) Standard specimen clamp with adapter 40 x 40 (silver) Fixture for specimen clamps, directional, silver Fixture for specimen clamps, rigid, silver Fixture for specimen clamps, directional, with 2 zero point indicators, silver Quick clamping system, complete Round specimen holder with adapter and 3 clamping rings, silver V-insert for Standard specimen clamp, silver Foil clamp - type 1, black Special wrench for EM specimen holder EM specimen holder fixture EM universal specimen holder EM flat specimen holder Megaclamp assembly RM22xx, silver RM CoolClamp Magnifying lens assembly Universal microscope carrier Fiber-optic light guide Light guide, gooseneck Instructions for Use V 2.1 RevF 11/2015

65 6. Optional Accessories Cold light sources Leica CLS 100X, V/50-60 Hz Leica CLS 100X, 230 V/50-60 Hz Leica CLS 100X, 240 V/50-60 Hz Backlighting assembly External power supply assembly Tray Cooling device assembly Section waste tray, large Section waste tray, small "Leica" brush w/magnet Demo Box RM22xx series Leica RM

66 7. Troubleshooting In the following table there is a list of the most common problems which can arise while working with the instrument, along with possible causes and troubleshooting procedures. Problem Possible cause Corrective action 7.1 Possible faults 1. Thick/thin sections The sections alternate between being thick and thin, or there is chattering in the sections, or the specimen is torn out of the embedding. In extreme cases, there are no sections whatsoever. The blade, knife holder or orientation is not clamped properly. The specimen is not clamped properly. Reclamp the blade, knife holders or orientation. Check that the cassette is securely clamped in the universal cassette clamp. If the universal cassette clamp is soiled by wax, clean the universal cassette clamp (refer to Chapter 8.1 "Cleaning and maintenance Universal Cassette Clamp"). When using cassettes whose lid is molded on, check that the broken edge allows the cassette to be clamped securely; if necessary, remove burrs or clamp the cassette into the universal cassette clamp horizontally instead of vertically. If the cassette dimensions are within the specified tolerances and the cassette still cannot be securely clamped into place, it may be that the universal cassette clamp is configured incorrectly or is defective. In this case, have Technical Service inspect and reconfigure the universal cassette clamp. 64 Instructions for Use V 2.1 RevF 11/2015

67 7. Troubleshooting Problem Possible cause Corrective action 2. Section compression The sections are very compressed, show folds or are squeezed together. 3. "Stripes" in sections For knife holder E The blade is dull. The pressure plate is damaged or incorrectly adjusted. Clearance angle of the knife/ blade too small. The blade is dull. The specimen is too warm. The sectioning speed is too fast. There is an accumulation of paraffin on the rear pressure plate of the knife holder. When using cassettes, particularly thin-walled cassettes, from a company other than Leica or Surgipath, the cassette may become deformed or other clamping problems may arise. If while trying to clamp the cassette you realize that it is not securely clamped into place, an alternative tensioning clamp must be used. Laterally displace the knife holder or insert a new blade. Insert a new pressure plate or use a new knife holder. Readjust the pressure plate. Systematically try several clearance angle adjustments, until the optimum angle is found. Use another area of the blade or a new blade. Cool the specimen before sectioning. Reduce the sectioning speed. Remove paraffin from this area on a regular basis. Leica RM

68 7. Troubleshooting Problem Possible cause Corrective action 4. Noises during sectioning The knife "sings" when sectioning hard specimens. The sections show scratches or chatter marks. 7.2 Instrument malfunctions 1. There is no further feed motion and thus no sectioning. The sectioning speed is too fast. The clearance angle is too wide. Insufficient clamping of specimen and/or knife holder. The front end position has been reached. 2. High blade consumption Too great of a sectioning force was applied. Turn the handwheel at a slower speed. Systematically decrease the clearance angle until the optimum adjustment is obtained. Check all screw and clamp connections on the specimen holder system and the knife holder. If necessary, tighten the levers and screws. Run the specimen back by turning the coarse driving wheel. Adjust the sectioning speed and/ or section thickness when trimming. Select a smaller section thickness, rotate the handwheel more slowly. 66 Instructions for Use V 2.1 RevF 11/2015

69 8. Cleaning and Maintenance 8.1 Cleaning the instrument Always remove the knife / blade before detaching the knife holder from the instrument. Always put the knives back into the knife case when not in use! Never place a knife anywhere with the cutting edge facing upwards and never try to catch a falling knife! When using cleaning agents, observe the manufacturer's safety instructions and the laboratory regulations valid in the country of use. When cleaning the outer surfaces, do not use xylene, scouring powders or solvents containing acetone or xylene. Xylene or acetone will damage the finished surfaces! Ensure that liquids do not enter the interior of the instrument during cleaning! Before each cleaning, carry out the following preparatory steps: Raise the specimen clamp to the upper end position and activate the handwheel lock. Switch the unit off and unplug it. Remove the blade from the knife holder and insert it in the receptacle at the bottom of the dispenser, or remove the knife from the knife holder and put it back in the knife case. Remove knife holder base and knife holder for cleaning. Remove the specimen from the specimen clamp. Remove section waste with a dry brush. Remove specimen clamp and clean separately. Instrument and outside surfaces If necessary, the varnished outside surfaces of the control panels can be cleaned with a mild commercial household cleaner or soap water and then be wiped with a moist cloth. To remove paraffin residue, xylene substitutes, paraffin oil, or paraffin removers such as "Para Gard" (Polysciences) can be used. The instrument must be completely dry before it can be used again. Always wipe the knife from the back of the knife to the cutting edge. NEVER wipe in the opposite direction - risk of injury! Clean using an alcohol-based solution or acetone. Leica RM

70 8. Cleaning and Maintenance Knife holder E Fig. 92 Take the knife holder apart for cleaning. To do so, proceed as follows: Fold knife guard (9) downward. Rotate the clamping lever (11) of the lateral displacement forwards and pull it out sideways. Push the knife holder base plate (86) with the pressure plate (83) until it can be removed from the segment arch (87). Rotate the blade clamping lever (10) downward and pull it out sideways. Remove pressure plate (83). Clean all parts of the knife holder. 11 If several knife holders are cleaned at the same time, the parts must NOT be mixed up! Failure to adhere to this may result in sectioning problems! For cleaning and removal of paraffin, do not use xylene or cleaning fluids containing alcohol (e.g. glass cleaner) Lay the removed parts on an absorbent cloth into the drying chamber (up to a max. of 65 C) and let the paraffin contamination run off. There is a burn hazard when removing the parts from the drying chamber (65 C). Wearing safety gloves is recommended! Dry knife holder and reassemble. After cleaning the moving parts, apply a thin coat of drive part oil to them (refer also to Chapter 8.3). When installing the pressure plate (83) make sure that the upper edge is parallel and level with the rear edge of the knife holder base plate (86). Fig Instructions for Use V 2.1 RevF 11/2015

71 Universal cassette clamp Detach cassette clamp (13) for a thorough cleaning, removing all paraffin residues. For cleaning, do not use xylene. Use xylene substitutes or paraffin removers such as "Para Gard." The cassette clamp (13) can also be placed in an oven heated to a maximum of 65 C, until the liquid wax escapes. Remove paraffin residues with a dry cloth. After such a cleaning procedure in an oven, always be sure to lubricate 91 the spiral springs (91) of the tension lever (60) (refer also to Chap. 8.3) and move it back and forth several times. Fig Maintenance instructions Only authorized and qualified service personnel may access the internal components of the instrument for service and repair! The instrument is basically maintenance-free. To ensure trouble-free operation of the instrument over a long period of time, the following is recommended by Leica: 1. Thoroughly clean the instrument on a daily basis. 2. Enter into a service contract at the latest at the end of the warranty period. For more information, please contact your local Leica technical service center. 3. Have the instrument inspected on a regular basis by a qualified service technician authorized by Leica. The intervals depend on how heavily the instrument is used. We recommend the following inspection intervals depending on the general workload of the instrument as defined in the two categories listed in the table: Category I Category II Number of sections per day: > 8,000 sections per day < 8,000 sections per day Workload (hours per day): > 5 hours per day < 5 hours per day Sectioning speed: Specimen material: Predominantly high sectioning speed Working with soft and hard specimens 8. Cleaning and Maintenance Slow to medium sectioning speed Predominantly soft specimen material Maintenance: Every 12 months Every 24 months Leica RM

72 8. Cleaning and Maintenance 8.3 Lubricating the instrument 95 Once a month, lubricate the following parts with the included drive parts oil No. 405 (1-2 drops are enough). 96 Instrument and specimen holder: The clamping key (95) of the quick clamping system. The T-piece (55) on the microtome base plate Guide rails (96) for the knife holder base on the microtome base plate. The T-piece on the knife holder base (56). 56 Knife holder E: Clamping lever (11) for the lateral displacement. T-piece (88) and guide (97) of the knife holder for the lateral displacement. 97 Clamping lever (10) for the blade. 8 Knife holder NZ Knife holders N and NZ: Sliding surfaces of the finger guard (8). Knurled nuts (73) for measuring height adjustment. Knife holder N 73 Fig Instructions for Use V 2.1 RevF 11/2015